Invitrogen

Phospho-ATF2 (Thr71) Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell Treatment to ensure that the antibody binds to the antigen stated. View Details

View all (40) ATF2 antibodies

Datasheet

Datasheet

Product Image Gallery

Phospho-ATF2 (Thr71) Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Phospho-ATF-2 pThr71 showing staining in the nucleus of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Anti- Phospho-ATF-2 pThr71 Polyclonal Antibody (Product # 44-295G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Phospho-ATF2 (Thr71) Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), U-87 MG treated with 25 µg/mL anisomycin for 30 minutes (Lane 2), K562 (Lane 3), K562 treated with 25 µg/mL anisomycin for 30 minutes (Lane 4). The blots were probed with Anti-Phospho-ATF-2 pThr71 Rabbit Polyclonal Antibody (Product # 44-295G, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). A ~ 54 kDa band corresponding to Phospho-ATF-2 pThr71 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Phospho-ATF2 (Thr71) Antibody in Cell Treatment — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot using Phospho-ATF2 (Thr71) Polyclonal Antibody (Product # 44-295G), shows increased levels of phospho-ATF2 (Thr71) in U-87MG and K562 cell lines upon Anisomycin treatment. {TM}

Product Details

Tested Applications

Dilution

Immunohistochemistry (Paraffin) (IHC (P))

1:20

Western Blot (WB)

1:1000

Product Specifications

Species Reactivity

Human

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Chemically synthesized phosphopeptide derived from a region of human ATF2 that contains threonine 71

Conjugate

Unconjugated

Form

Liquid

Purification

Antigen affinity chromatography

Storage buffer

Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/mL BSA

Contains

0.05% sodium azide

Storage conditions

-20°C

RRID

AB_2533624

Product Specific Information

This antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non phosphorylated ATF2. The final product is generated by affinity chromatography using an ATF2-derived peptide phosphorylated at threonine 71. The peptide sequence is conserved in mouse, rat, chicken and frog. Suggested Western blot positive controls: NIH3T3 + anisomycin, or Jurkat + 10 µg/mL anisomycin for 60 minutes.

Target Information

ATF-2 is a member of the group of bZip transcription factors. Heterodimer formation between members of the bZip group is common and is believed to add diversity to the cis-acting elements at which binding of the dimers is directed. Specifically, ATF-2 may dimerize with c-Jun, as occurs in response to E1a, and in so doing shift the binding preference of c-Jun toward ATF/CRE sites (1-3). Deletion analysis has indicated that the N-terminal region of ATF-2 containing threonine at residues 69 and 71 are essential for this purpose. These threonine residues are phosphorylated by JNK/SAPK for transcriptional activation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Activating 2; Activating transcription factor 2; activating transcription factor 2 splice variant ATF2-var2; ATF2; cAMP response element binding protein CRE- BP1; cAMP response element-binding protein CRE-BP1; cAMP responsive element binding protein 2, formerly; cAMP-dependent transcription factor ATF-2; CREB-2; CREB2; CREBP1; Cyclic AMP-dependent transcription factor ATF-2; Cyclic AMP-responsive element-binding protein 2; Cyclic-AMP-dependent ATF-2; HB16; Histone acetyltransferase ATF2

Gene Aliases: ATF2; CRE-BP1; CREB-2; CREB2; CREBP1; HB16; TREB7

UniProt ID: (Human) P15336

Entrez Gene ID: (Human) 1386

Molecular Function: nucleic acid binding transcription factor